Frozen vs paraffin: Confocal microscopy - (Feb/09/2012 )

Hi everyone!

I was wondering if anyone knows which tissue preperation (frozen vs paraffin) is better for confocal microscopy. Also, I will be using antibodies against phospho-proteins and I have heard that these do no do so well when tissue has been embedded in paraffin.

Any insight would be greatly appreciated!
Thanks,
Carmen

WIth paraffin, you can get thin sections. and wIth frozen you get thicker sections. Thinner sections are good for staining.
I have no experience comparing phospho antibodies on sections.

Have you seen this thread ? There's also something on anti-phospho antibodies...
http://www.protocol-online.org/biology-forums-2/posts/16990.html

Regarding the confocal, either frozen or paraffin-embedded sections should work as long as the antibody works. Because of how the confocal microscope works, it is able to see through the tissue section to a single plane, so the thickness of the frozen sections doesn't really affect the confocal microscope like it would in a normal fluorescent microscope. So if you find that you get better staining/morphology with the frozen sections then use the frozen sections.